A variety of Clostridium sp. strains which secrete toxins having neurotoxic effects have been discovered since the 1890s up to the present time, and the characterization of toxins that are secreted from these strains has been made over the past 70 years (Schant, E. J. et al., Microbiol. Rev., 56:80, 1992). Among these toxins, botulinum toxin inhibits the exocytosis of acetylcholine at the cholinergic presynapse of a neuromuscular junction in animals having neurological function to thereby cause asthenia. Thus, efforts have recently been made to use the neurotoxicity of botulinum toxin for cosmetic or therapeutic purposes. Technologies for using botulinum toxin for treatment of optic diseases (U.S. Pat. No. 6,265,379), pain (U.S. Pat. No. 6,113,915), various autonomic nerve disorders, including sweat gland disorders (U.S. Pat. No. 5,766,605), migraine headache pain (U.S. Pat. No. 5,714,468), post-operative pain and visceral pain (U.S. Pat. No. 6,464,986), psoriasis and dermatitis (U.S. Pat. No. 5,670,484), various cancers (U.S. Pat. Nos. 6,139,845 and 6,063,768), and neurogenic inflammation (U.S. Pat. No. 6,063,768), etc. have been proposed or attempted. However, botulinum toxin, a protein agent, has a problem in that it is not easy to formulate into pharmaceutical compositions and is also not easy to store, distribute and manage. This is attributable to the instability of the protein, and the problem is serious in the case of protein agents such as botulinum toxin, which are formulated into pharmaceutical compositions at a very low concentration. Botulinum toxin protein has the property of adhering to a solid surface, and for this reason, when the protein is injected into a container, a portion of the protein may adhere to the inner wall of the container to cause the loss of the active ingredient, and the protein may be easily oxidized or degraded into small fragments. For this reason, in order to prevent the denaturation of botulinum toxin to the greatest possible extent, botulinum toxin purified in a production process thereof is distributed as freeze-dried powder, which is diluted in a saline immediately before use in clinical applications and administered to patients in the form of liquid. However, in this case, there is also a problem in that medical accidents are highly likely to occur due to human errors such as a dilution factor error caused by the user or contamination of a dilution saline.
Therefore, it is urgently needed to develop stabilizers that can prevent protein denaturation even during the production and distribution of a liquid formulation of botulinum toxin.
In the prior art, albumin was actively used as a stabilizer to maintain the activity of botulinum toxin. However, due to the risk of cross infection and side effects of animal-derived components, the development of non-animal formulations has recently been required. In response to this requirement, US Patent Application Publication No. 2007-0134199 discloses a botulinum toxin composition comprising either glutamine and glutamic acid or asparagine and aspartic acid as amino acids, and Korean Patent No. 1087017 discloses a botulinum toxin composition comprising methionine as a stabilizer. However, these patent documents do not suggest remarkable effects under suitable conditions according to the temperature and pH of the human body.
Therefore, the present invention is directed to a liquid formulation containing botulinum toxin and stabilizing agent, and preparation method therefor. A pharmaceutical composition comprising botulinum toxin according to the present invention may contain arginine, glutamic acid, or aspartic acid as a stabilizer, or may contain gluconolactone buffer, or tartaric acid buffer as a stabilization buffer for botulinum toxin. AND it was proved a significant effect on the stabilization of botulinum toxin under suitable conditions according to the temperature and pH of the human body. Thus, it is expected that the pharmaceutical composition of the present invention will greatly contribute to the safe and convenient medical use of botulinum toxin.